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Image Search Results
Figure S2 . " width="100%" height="100%">
Journal: Cell Reports
Article Title: Mechanistic insights into COVID-19 by global analysis of the SARS-CoV-2 3CL pro substrate degradome
doi: 10.1016/j.celrep.2021.109892
Figure Lengend Snippet: Characterization of 3CL pro cleavage specificity (A and B) MALDI-TOF-MS spectra of synthetic peptides spanning P4–P4’ of protein cleavage sites after incubation with 3CL pro (1:20 molar ratio, E:S). Product generation (red) and substrate consumption (black) were calculated as the peak area normalized to the total peak area in the spectrum. Apparent (app) k cat / K M values for 1 μM 3CL pro to convert 50% of substrate in 5, 15, 30, 60, 120 or 240 min are listed alongside bins of 4 peptides that share similar kinetic values arranged on a row-by-row basis. P4–P4’ sequence alignment using the Shapley color scale. Green protein names had cut sites identified by Edman sequencing of recombinant substrate digests. Boxed peptides, no cleavage. (C–I) Structures of the highest-ranked of 50,000 models of the active site of 3CL pro protomer 1 (PDB: 6XHM ) docked with P4–P4’ peptides from six 3CL pro substrates exhibiting a range of app k cat / K M values (circled in B). (C) 3CL pro dimer. Protomer 1, gray surface or green ribbons with catalytic Cys 145 shown. Protomer 2, orange surface with Ser 1 shown. Docking models with P4–P4’ peptide of: (C and D) RPAP1 (I_sc = −31.7), (E) IMA4 (I_sc = −30.6), (F) PTBP1 (I_sc = −31.7), (G) RBM15 (I_sc = −39.65), (H) MAP4K5 (I_sc = −28.1), and (I) CREB1 (I_sc = −30.3). Blue and red sticks, P and P’ amino acid residues, respectively. Yellow dashed sticks, hydrogen-bonds. See also
Article Snippet: The primary antibodies and dilutions used were: mouse monoclonal anti-SARS-CoV-2 nucleocapsid antibody (1:1,000, Invitrogen, MA5-29981, RRID: AB_2785780 );
Techniques: Incubation, Sequencing, Recombinant
Figure 1 " width="100%" height="100%">
Journal: Cell Reports
Article Title: Mechanistic insights into COVID-19 by global analysis of the SARS-CoV-2 3CL pro substrate degradome
doi: 10.1016/j.celrep.2021.109892
Figure Lengend Snippet: 3CL pro cleavage sites and substrate proteins stringently identified in human embryonic kidney (HEK293) and human lung epithelial (BEAS-2B) cells, related to
Article Snippet: The primary antibodies and dilutions used were: mouse monoclonal anti-SARS-CoV-2 nucleocapsid antibody (1:1,000, Invitrogen, MA5-29981, RRID: AB_2785780 );
Techniques:
Figure S1 and , , , Journal: Cell Reports
Article Title: Mechanistic insights into COVID-19 by global analysis of the SARS-CoV-2 3CL pro substrate degradome
doi: 10.1016/j.celrep.2021.109892
Figure Lengend Snippet: 3CL pro cleavage sites in human proteins identified by TAILS (A) Experimental design. Neo-N-termini of 3CL pro -cleaved substrates in HEK293 and BEAS-2B human lung epithelial cell lysates were isolated and identified by TAILS LC-MS/MS. Only those neo-N-termini in ≥ 2/3 HEK293 or ≥ 7/9 BEAS-2B independent cell experiments were considered for further substrate winnowing. (B) Classification of 1,649 quantified N-termini in N = 12 independent experiments. (C) 3CL pro candidate substrate cleavage site specificities (n = 292) versus other quantified neo-N-termini (n = 663). (D) Cellular distribution of high confidence cleavage sites (n = 102) in 101 human substrates after substrate winnowing . (E) Substrate Reactome gene set enrichment by hypergeometric distribution followed by FDR correction. Node radius designates gene enrichment; line widths are proportional to the overlap of shared substrates between connected nodes sharing ≥ 20% genes. See also
Article Snippet: The primary antibodies and dilutions used were: mouse monoclonal anti-SARS-CoV-2 nucleocapsid antibody (1:1,000, Invitrogen, MA5-29981, RRID: AB_2785780 );
Techniques: Isolation, Liquid Chromatography with Mass Spectroscopy, Comparison, Control
Journal: Cell Reports
Article Title: Mechanistic insights into COVID-19 by global analysis of the SARS-CoV-2 3CL pro substrate degradome
doi: 10.1016/j.celrep.2021.109892
Figure Lengend Snippet: 3CL pro cleaves RPAP1 and PTBP1, altering PTBP1 localization (A) Locations of 3CL pro cleavage sites in RPAP1 and PTBP1 identified by TAILS neo-N-terminal peptides (red) and Edman sequencing (green). Representative MS/MS spectra of cleaved neo-N-terminal peptides. (B) SDS-PAGE and Edman sequencing of human recombinant RPAP1 and PTBP1 incubated with 3CL pro +/− inhibitor GC376, or 3CL pro -C145A (1:5 mol/mol, E:S). Δ-substrate, no sequence obtained. (C) MALDI-TOF-MS kinetic analyses of 3CL pro cleavage of synthetic P4–P4’ peptides. (D) RPAP1 and PTBP1 immunoblots of primary HAECs lysates from 5 donors incubated with 3CL pro or 3CL pro -C145A (1:200 w/w, E:S) for 18 h, 37°C. (E and F) (E) Immunoblots of 3CL pro , N-protein and (F) PTBP1 of Vero E6 cells infected with SARS-CoV-2 at a MOI 0.1 for 24 (n = 4) and 48 (n = 4) hpi, or mock (n = 3) ∗ , unspecific bands. (G) Subcellular localization of PTBP1 by confocal imaging. SARS-CoV-2-infected Vero E6 cells (N = 5, scale bar 50 μm). Cyan boxes, Spike-negative (S–) uninfected cells. Green boxes, Spike-positive (S+) infected cells. Enlarged detail of mock-infected, S+ and S– cells in the same field, scale bar 10 μm. (H) Nuclear to cytoplasmic ratio of PTBP1 was quantified for mock and Spike-positive SARS-CoV-2 infected cells. Statistical significance was assessed by Student’s t test (mean ± SD, N = 5, n = 51 cells, ∗ p ≤ 0.05). β-actin and β-tubulin loading controls. See also .
Article Snippet: The primary antibodies and dilutions used were: mouse monoclonal anti-SARS-CoV-2 nucleocapsid antibody (1:1,000, Invitrogen, MA5-29981, RRID: AB_2785780 );
Techniques: Sequencing, Tandem Mass Spectroscopy, SDS Page, Recombinant, Incubation, Western Blot, Infection, Imaging
Journal: Cell Reports
Article Title: Mechanistic insights into COVID-19 by global analysis of the SARS-CoV-2 3CL pro substrate degradome
doi: 10.1016/j.celrep.2021.109892
Figure Lengend Snippet: Hippo pathway substrate validation (A) 3CL pro cleavage sites in MAP4K5 and CREB1 identified by TAILS neo-N-terminal peptides (red) and Edman sequencing (green). Representative MS/MS spectra of cleaved neo-N-terminal peptides. (B) MALDI-TOF-MS kinetic analyses of 3CL pro cleavage of P4–P4’ peptides of MAP4K5 and CREB1. (C) SDS-PAGE, Edman sequencing (green) and immunoblot validation of human MAP4K5 and CREB1 substrates incubated with 3CL pro +/− inhibitor GC376, or 3CL pro -C145A (1:5 mol/mol, E:S). ΔMAP4K5 or ΔCREB1, no sequence obtained. (D and H) (D) YAP1, MAP4K5, CREB1 and (H) FYCO1 and FAF1 immunoblots of lysates from primary HAECs incubated with 3CL pro or 3CL pro -C145A (1:200 w/w, E:S) for 18 h, 37°C. (E and F) (E) YAP1 and (F) MAP4K5 immunoblots of Vero E6 cells at 24 (n = 4) and 48 (n = 4) hpi (MOI 0.1) or mock (n = 3). MAP4K5 activity assay measured as ATP consumption using myelin basic protein as substrate. The area under the curve was calculated and compared by Student’s t test, (mean ± SD, n = 2, ∗∗ p ≤ 0.01). (G, I, and J) (G) Lysates of human Calu-3 lung cells infected with SARS-CoV-2 (MOI 0.1 and 1.0, n = 4, mock n = 2) were immunoblotted for (G) CREB1 48 hpi, (I) FYCO1 24 hpi and (J) FAF1 48 hpi. Statistical analysis of the relative amount of full-length protein (E and F) or proteolytic bands (G, I, and J) identified by molecular weights relative to β-actin was assessed by one-way ANOVA and Dunnett’s multiple comparisons test. Box and whiskers (min to max) plots, ∗∗∗ p ≤ 0.001, ∗∗ p ≤ 0.01, ∗ p ≤ 0.05, ns p > 0.05. β-actin and β-tubulin loading controls. See also and .
Article Snippet: The primary antibodies and dilutions used were: mouse monoclonal anti-SARS-CoV-2 nucleocapsid antibody (1:1,000, Invitrogen, MA5-29981, RRID: AB_2785780 );
Techniques: Biomarker Discovery, Sequencing, Tandem Mass Spectroscopy, SDS Page, Western Blot, Incubation, Activity Assay, Infection
Figure S6 . " width="100%" height="100%">
Journal: Cell Reports
Article Title: Mechanistic insights into COVID-19 by global analysis of the SARS-CoV-2 3CL pro substrate degradome
doi: 10.1016/j.celrep.2021.109892
Figure Lengend Snippet: 3CL pro disrupts galectin-8 binding to Spike in antiviral-autophagy (A) Immunoblot of human galectin-8 (Gal8) in BEAS-2B cells in response to IFN- α , IFN-β, or vehicle. One way ANOVA and Dunnett's posthoc test (mean ± SD, n = 3 each, *** p ≤ 0.001, * p ≤ 0.05). (B) MALDI-TOF-MS of intact versus 3CL pro -cleaved synthetic Gal8 P4–P4’ peptide. (C) SDS-PAGE and Edman sequencing of Gal8 incubated with 3CL pro +/− inhibitor GC376, or 3CL pro C145A (1:5 mol/mol, E:S). (D) Structural model of Gal8 docked onto 3CL pro . 3CL pro cleavage site identified by the neo-N-terminal peptide (red) in 9/9 independent TAILS analyses. (E) Gal8 immunoblots of lysates from primary HAECs incubated with 3CL pro or 3CL pro -C145A (1:200 w/w, E:S) for 18 h, 37°C (N = 5). (F) Gal8 immunoblot of infected Calu-3 cells at 24 (n = 4) and 48 (n = 4) hpi (MOI 1.0, mock n = 3). β-actin and β-tubulin loading controls. (G) ELISA of SARS-CoV-2 Spike S1 protein binding intact Gal8 or 3CL pro -cleaved (ΔGal8) (mean ± SD, n = 2, N = 2, ∗∗∗∗ p ≤ 0.0001, ∗∗ p ≤ 0.01, ns p > 0.05, two-way ANOVA with Šídák’s multiple comparison test). (H) Immunoprecipitation (IP) by α-FLAG agarose-beads of HeLa cell lysates co-transfected with GFP-NDP52, WT-Gal8-FLAG or C-Gal8 (159-317)-FLAG. (I) Intact or cleaved Gal8 binding immobilized NDP52 detected with anti-C-Gal8 antibody. Student's t test, (mean ± SD, n = 3, N = 2, ns p > 0.05). (J) NDP52 binding Spike S1-associated intact or cleaved ΔGal8 (mean ± SD, n = 2, N = 2, ∗∗∗∗ p ≤ 0.0001, ∗∗∗ p ≤ 0.001, ∗ p ≤ 0.05, ns p > 0.05, two-way ANOVA with Šídák’s multiple comparison test). (K) Confocal microscopy of NDP52 and FLAG immunofluorescence after osmotic shock of HEK293 cells transfected with FLAG-tagged (WT) Gal8, N-Gal8 (1-158), or C-Gal8 (159-317). Scale bar, 20 μm. Quantification of NDP52 and Gal8-positive puncta, mean ± SD, n = 30, N = 5, ∗∗∗∗ p ≤ 0.0001, ns p > 0.05, one-way ANOVA with Tukey’s multiple comparison test. (L) Human lung tissue sections from normal subjects (N = 3) and post-mortem COVID-19 patients (N = 4) stained with hematoxylin and eosin (H&E), DAPI (blue) and immunofluorescence on the same sections for Gal8 (green), NDP52 (red), Spike (magenta) and merged image (orange). Scale bar, 50 μm. (M) Numbers of immunopositive cells for Gal8, NDP52 and Gal8 colocalized with NDP52 in healthy versus COVID-19 lung tissue. Total cell count distribution shown as a scatter dot plot, bar = median. See also
Article Snippet: The primary antibodies and dilutions used were: mouse monoclonal anti-SARS-CoV-2 nucleocapsid antibody (1:1,000, Invitrogen, MA5-29981, RRID: AB_2785780 );
Techniques: Binding Assay, Western Blot, SDS Page, Sequencing, Incubation, Infection, Enzyme-linked Immunosorbent Assay, Protein Binding, Comparison, Immunoprecipitation, Transfection, Confocal Microscopy, Immunofluorescence, Staining, Cell Counting
Journal: Cell Reports
Article Title: Mechanistic insights into COVID-19 by global analysis of the SARS-CoV-2 3CL pro substrate degradome
doi: 10.1016/j.celrep.2021.109892
Figure Lengend Snippet: Protein-protein interaction landscape of 3CL pro cell substrates (A) Network of direct (solid line) and physically associated (dashed line) interactors (blue dots) of the 101 3CL pro substrates (red circles) generated by the Intact-database (accessed May 2021). Interactors of ≥ 2 substrates are in magenta, only interactors of ≥ 3 substrates are labeled, the number of substrate interactions is indicated by circle size. Orange circles, candidate substrates manually annotated. (B) Top 5 CORUM protein complexes statistically enriched in the 3CL pro substrate-human interactome. (C) One-step direct neighborhood protein-protein interaction network of 3CL pro human substrates (n = 74, red circles) that are directly connected to SARS-CoV-2 proteins (n = 26, yellow diamonds) or connected via direct neighbors (n = 197). Assembled from IMex/Intact Coronavirus dataset accessed June 2, 2021. Yellow diamonds, viral proteins. Blue circles, direct interactors connecting viral proteins to substrates. Magenta circles, direct interactors connecting viral proteins to ≥ 2 substrates. Black edge, SARS-CoV-2/human protein interactions. See also .
Article Snippet: The primary antibodies and dilutions used were: mouse monoclonal anti-SARS-CoV-2 nucleocapsid antibody (1:1,000, Invitrogen, MA5-29981, RRID: AB_2785780 );
Techniques: Generated, Labeling
Journal: Cell Reports
Article Title: Mechanistic insights into COVID-19 by global analysis of the SARS-CoV-2 3CL pro substrate degradome
doi: 10.1016/j.celrep.2021.109892
Figure Lengend Snippet:
Article Snippet: The primary antibodies and dilutions used were: mouse monoclonal anti-SARS-CoV-2 nucleocapsid antibody (1:1,000, Invitrogen, MA5-29981, RRID: AB_2785780 );
Techniques: Virus, Recombinant, Sequencing, Fluorescence, Modification, Protease Inhibitor, Polymer, Staining, Blocking Assay, Binding Assay, Activity Assay, Western Blot, Synthesized, Software, Control, Targeted Proteomics, Microscopy, Spectrophotometry, Mass Spectrometry
Journal: ACS Omega
Article Title: Inhibition of the 3CL Protease and SARS-CoV-2 Replication by Dalcetrapib
doi: 10.1021/acsomega.1c01797
Figure Lengend Snippet: Docking of dalcetrapib-thiol in the active site of 3CLpro of the SARS-CoV-2. (A) Docking of dalcetrapib-thiol (labeled as dalcetrapib-thiol) in 3CLpro (PDB 6W63) using the standard protocol, which resulted in a distance of 3.6 Å between the thiol of dalcetrapib-thiol and Cys145. Dalcetrapib-thiol (carbon in magenta) and the surrounding residues (carbon in green) are shown in stick mode. The hydrogen bond between Dalcetrapib-thiol and the main chain amide of Glu166 is shown as a black dashed line. (B) Surface representation of (A) with the S1′, S1, S2, and S4 subsites indicated. (C) Docking of dalcetrapib-thiol to the active site of 3CLpro (PDB 6W63) using the SCAR protocol, which resulted in a distance of 2.0 Å between the thiol of dalcetrapib-thiol and Cys145. Dalcetrapib-thiol (carbon atoms in cyan) and the surrounding residues (carbon atoms in green) are shown in stick mode. (D) Surface representation of (C) with the S1′, S1, S2, and S4 subsites indicated.
Article Snippet:
Techniques: Labeling
Journal: ACS Omega
Article Title: Inhibition of the 3CL Protease and SARS-CoV-2 Replication by Dalcetrapib
doi: 10.1021/acsomega.1c01797
Figure Lengend Snippet: (A) Dalcetrapib directly inhibits SARS-CoV-2 3CLpro activity. The IC 50 for dalcetrapib was determined by measuring fluorescence released from the quenched peptide exposed to the protease. Dalcetrapib was first de-esterified by basic hydrolysis of the ester bond to obtain dalcetrapib-thiol, which was then preincubated with 3CLpro for 24 h before a further 30 min preincubation in the presence of 1 mM DTT. Slopes of the relative fluorescence unit (RFU) increase were derived after the first 15 min of incubation with the peptide. Curve-fitting was performed using a GraphPad Prism four-parameter method. (B) Procedure for the assessment of the reversibility of 3CLpro inhibition by dalcetrapib. 3CLpro was incubated for 24 h with a buffer (control) or dalcetrapib-thiol produced as indicated in the method section. The incubation mixtures were then diluted or not 1000-fold through three ultrafiltration cycles to remove free dalcetrapib-thiol and concentrate 3CLpro. Samples labeled #1, #2, #3, and #4 were analyzed for 3CLpro activity (shown in panel C). (C) Assessment of the reversibility of 3CLpro inhibition by dalcetrapib. Protease activity “before” and “after” repeated dilution/filtration cycles. Dalcetrapib-thiol was preincubated with 3CLpro for 24 h at room temperature in the presence of 1 mM DTT before addition of the quenched peptide substrate. Slopes of the relative fluorescence unit (RFU) increase were derived after the first 15 min of incubation with the peptide.
Article Snippet:
Techniques: Activity Assay, Fluorescence, Derivative Assay, Incubation, Inhibition, Control, Produced, Labeling, Filtration
Journal: Nature
Article Title: Multiple pathways for SARS-CoV-2 resistance to nirmatrelvir
doi: 10.1038/s41586-022-05514-2
Figure Lengend Snippet: a , Changes in IC 50 during passaging of SARS-CoV-2 with nirmatrelvir. Vero E6 cells were infected in triplicate with SARS-CoV-2 (USA-WA1/2020) and passaged to fresh cells every 3 days for 30 passages . b – d , Validation of nirmatrelvir resistance for the indicated passage from each of the three lineages (A ( b ), B ( c ) and C ( d )). e , Inhibition of passage 30 viruses from each lineage by remdesivir. f , Mutations in 3CL pro found in the indicated passages from each lineage. Dots indicate WT at that residue. Mutations are shaded according to frequency. g , Residues mutated with passaging in Vero E6 cells overlaid onto the 3CL pro structure with nirmatrelvir bound. The Cα of each mutated residue is denoted by a red sphere. The 3CL pro –nirmatrelvir complex was downloaded from PDB under accession code 7VH8. a – e , Error bars denote mean ± s.e.m of four technical replicates.
Article Snippet: Extended Data Fig. 5 Individual inhibition curves of recombinant live SARS-CoV-2 carrying single and
Techniques: Passaging, Infection, Inhibition
Journal: Nature
Article Title: Multiple pathways for SARS-CoV-2 resistance to nirmatrelvir
doi: 10.1038/s41586-022-05514-2
Figure Lengend Snippet: a , Passaging scheme: 480 wells were infected with SARS-CoV-2-mNeonGreen and passaged to fresh Huh7-ACE2 cells every 3–4 days, with the concentration of drug doubled every two passages. b , Validation of nirmatrelvir resistance of three wells from passage 16. These viral populations had the following mutations: 3A8 (T21I, T304I), 1E11 (T21I, N51Y, T304I) and 5A2 (L50F, E166V). See Supplementary Table for exact frequencies. Representative curves from a single experiment from two biologically independent experiments are shown. Error bars denote mean ± s.e.m of three technical replicates. c , Mutations in 3CL pro found in passage 16 from 53 wells. Dots indicate WT at that residue. Mutations are shaded according to frequency. d , Residues mutated in passaging in Huh7-ACE2 cells overlaid onto the 3CL pro structure with nirmatrelvir bound. All 23 mutated residues across all resistant populations are indicated for any individual isolate having between one and six mutations. The Cα of each residue that was mutated is denoted by a red sphere for mutations observed more than ten times, and is denoted by an orange sphere for mutations observed fewer than ten times. The 3CL pro –nirmatrelvir complex was downloaded from PDB under accession code 7VH8.
Article Snippet: Extended Data Fig. 5 Individual inhibition curves of recombinant live SARS-CoV-2 carrying single and
Techniques: Passaging, Infection, Concentration Assay
Journal: Nature
Article Title: Multiple pathways for SARS-CoV-2 resistance to nirmatrelvir
doi: 10.1038/s41586-022-05514-2
Figure Lengend Snippet: Dots indicate wild-type at that cut site. Note that nsp4/5 M(P6’)I = M6I, nsp5/6 S(P6)P = S301P, and nsp5/6 T(P3)I = T304I in 3CL pro .
Article Snippet: Extended Data Fig. 5 Individual inhibition curves of recombinant live SARS-CoV-2 carrying single and
Techniques:
Journal: Nature
Article Title: Multiple pathways for SARS-CoV-2 resistance to nirmatrelvir
doi: 10.1038/s41586-022-05514-2
Figure Lengend Snippet: a , Phylogenetic tree of sequences from passaging in Huh7-ACE2 cells. Only sequences with mutations are shown. Sequences are denoted as passage number, followed by the well number. Mutations that arose along particular branches are annotated in red; ‘-’ denotes when a mutation appears to have been lost from a particular branch. b , Observed pathways for nirmatrelvir resistance in Huh7-ACE2 cells. The most commonly observed mutations in passage 16 were used to build these pathways ( and Supplementary Table ). Nodes are shaded from dark to light, with founder mutations darker. Percentages indicate the frequency by which child nodes derive from the immediate parental node. Descendent arrows that do not sum to 100% indicate that a proportion did not advance beyond the indicated mutations in the experiment. c , Growth assay with recombinant live SARS-CoV-2 carrying single (top) and combination 3CL pro mutations (bottom). Huh7-ACE2 cells were infected with 0.01 multiplicity of infection (MOI) of virus, and luminescence was quantified at the indicated time points. S144A, E166V and T21I + S144A are statistically significant from WT at 48 h (two-way analysis of variance with Geisser–Greenhouse correction followed by Dunnett’s multiple comparisons test; P = 0.0039, P = 0.0006, P = 0.0006, respectively). Representative curves from a single experiment from two biologically independent experiments are shown. Error bars denote mean ± s.e.m of three technical replicates. RLU, relative luminescence units.
Article Snippet: Extended Data Fig. 5 Individual inhibition curves of recombinant live SARS-CoV-2 carrying single and
Techniques: Passaging, Mutagenesis, Growth Assay, Recombinant, Infection
Journal: Nature
Article Title: Multiple pathways for SARS-CoV-2 resistance to nirmatrelvir
doi: 10.1038/s41586-022-05514-2
Figure Lengend Snippet: The Cα of each residue that was mutated is denoted with a red sphere. The 3CL pro -nirmatrelvir complex was downloaded from PDB under accession code 7VH8.
Article Snippet: Extended Data Fig. 5 Individual inhibition curves of recombinant live SARS-CoV-2 carrying single and
Techniques:
Journal: Nature
Article Title: Multiple pathways for SARS-CoV-2 resistance to nirmatrelvir
doi: 10.1038/s41586-022-05514-2
Figure Lengend Snippet: a , Individual inhibition curves of recombinant live SARS-CoV-2 carrying single (left) and combination 3CL pro mutations (right) by nirmatrelvir. Representative curves from a single experiment from three biologically independent experiments are shown. Error bars denote mean ± s.e.m of three technical replicates. b , Inhibition of recombinant live SARS-CoV-2 carrying single and combination 3CL pro mutations by nirmatrelvir, ensitrelvir and remdesivir. Values shown are fold change of mean values in IC 50 relative to inhibition of WT from three biologically independent experiments.
Article Snippet: Extended Data Fig. 5 Individual inhibition curves of recombinant live SARS-CoV-2 carrying single and
Techniques: Inhibition, Recombinant
Journal: Nature
Article Title: Multiple pathways for SARS-CoV-2 resistance to nirmatrelvir
doi: 10.1038/s41586-022-05514-2
Figure Lengend Snippet: a , Overlay of nirmatrelvir and ensitrelvir binding to 3CL pro . b , Several of the residues involved in direct interaction with nirmatrelvir. c , Several of the residues involved in formation of the S1 subsite. d , Interaction of L167 with nirmatrelvir. In a - d , nirmatrelvir is shown in yellow, enstirelvir is shown in lime green, the 3CL pro -nirmatrelvir complex is shown in marine, and the 3CL pro -ensitrelvir complex is shown in gray. Protomer A is shown in marine and protomer B is shown in green. Hydrogen bonds are indicated as black dashes. The 3CL pro -nirmatrelvir complex and 3CL pro -ensitrelvir complex were downloaded from PDB under accession codes 7VH8 and 7VU6, respectively.
Article Snippet: Extended Data Fig. 5 Individual inhibition curves of recombinant live SARS-CoV-2 carrying single and
Techniques: Binding Assay